• Title/Summary/Keyword: Morphology Control

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Induction of Angiogenesis by Matrigel Coating of VEGF-Loaded PEG/PCL-Based Hydrogel Scaffolds for hBMSC Transplantation

  • Jung, Yeon Joo;Kim, Kyung-Chul;Heo, Jun-Young;Jing, Kaipeng;Lee, Kyung Eun;Hwang, Jun Seok;Lim, Kyu;Jo, Deog-Yeon;Ahn, Jae Pyoung;Kim, Jin-Man;Huh, Kang Moo;Park, Jong-Il
    • Molecules and Cells
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    • v.38 no.7
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    • pp.663-668
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    • 2015
  • hBMSCs are multipotent cells that are useful for tissue regeneration to treat degenerative diseases and others for their differentiation ability into chondrocytes, osteoblasts, adipocytes, hepatocytes and neuronal cells. In this study, biodegradable elastic hydrogels consisting of hydrophilic poly(ethylene glycol) (PEG) and hydrophobic poly(${\varepsilon}$-caprolactone) (PCL) scaffolds were evaluated for tissue engineering because of its biocompatibility and the ability to control the release of bioactive peptides. The primary cultured cells from human bone marrow are confirmed as hBMSC by immunohistochemical analysis. Mesenchymal stem cell markers (collagen type I, fibronectin, CD54, $integrin1{\beta}$, and Hu protein) were shown to be positive, while hematopoietic stem cell markers (CD14 and CD45) were shown to be negative. Three different hydrogel scaffolds with different block compositions (PEG:PCL=6:14 and 14:6 by weight) were fabricated using the salt leaching method. The hBMSCs were expanded, seeded on the scaffolds, and cultured up to 8 days under static conditions in Iscove's Modified Dulbecco's Media (IMDM). The growth of MSCs cultured on the hydrogel with PEG/PCL= 6/14 was faster than that of the others. In addition, the morphology of MSCs seemed to be normal and no cytotoxicity was found. The coating of the vascular endothelial growth factor (VEGF) containing scaffold with Matrigel slowed down the release of VEGF in vitro and promoted the angiogenesis when transplanted into BALB/c nude mice. These results suggest that hBMSCs can be supported by a biode gradable hydrogel scaffold for effective cell growth, and enhance the angiogenesis by Matrigel coating.

Effect of Stem Cell Transplantation on Pain Behavior and Locomotor Function in Spinal Cord Contusion Model

  • Park, Hea-Woon;Kim, Su-Jeong;Cho, Yun-Woo;Hwang, Se-Jin;Lee, Won-Yub;Ahn, Sang-Ho;Jang, Sung-Ho
    • The Journal of Korean Physical Therapy
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    • v.22 no.3
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    • pp.79-85
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    • 2010
  • Purpose: Many trials for new therapeutic approaches such as stem cell-based transplantation have been conducted to improve the repair and regeneration of injured cord tissue and to restore functions following spinal cord injury (SCI) in animals and humans. Adipose tissue-derived stromal cells (ATSCs) have multi-lineage potential to differentiate into cells with neuron-like morphology. Most studies of stem cell transplantation therapy after SCI are focused on cellular regeneration and restoration of motor function, but not on unwanted effects after transplantation such as neuropathic pain. This study was focused on whether transplantation of ATSCs could facilitate or attenuate hindpaw pain responses to heat, cold and mechanical stimulation, as well as on improvement of locomotor function in a rat with SCI. Methods: A spinal cord injury rat model was produced using an NYU impactor by dropping a 10 g rod from a height of 25 mm on to the T9 segment. Human ATSCs (hATSCs; approximately $5{\times}10^5$ cells) or DMEM were injected into the perilesional area 9 days after the SCI. After transplantation, hindpaw withdrawal responses to heat, cold and mechanical allodynia were measured over 7 weeks. Motor recovery on the Basso, Beattie, and Bresnahan (BBB) locomotor rating scale and on the inclined plane test were also evaluated. Results: The present study demonstrated that increased hindpaw withdrawal responses to cold allodynia was observed in both groups after transplantation, but the development of cold-induced allodynia in the hATSC transplantation group was significantly larger than in the control group. The difference between the two groups in locomotor functional improvement after SCI was also significant. Conclusion: Careful consideration not only of optimal functional benefits but also of unintended side effects such as neuropathic pain is necessary before stem cell transplantation therapy after SCI.

Changes of Growth, Morphology and Microcystin Production in Microcystis aeruginosa in Response to Zooplankton Culture Media Filtrate (동물플랑크톤 배양여과액에 의한 Microcystis aeruginosa의 성장,형태 및 microcystin 생성량의 변화)

  • Ha, Kyong;Jang, Min-Ho;Jung, Jong-Mun;Joo, Gea-Jae
    • Korean Journal of Ecology and Environment
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    • v.36 no.1 s.102
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    • pp.1-8
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    • 2003
  • Growth, colony formation and microcystin production of 'low-toxic' Microcysits aeruginosa $K{\"{u}}tzing$ were examined in relation to the 'info-chemicals' released by zooplankton. Algae were cultured in a medium with or without filtered water taken from cultures of Daphnia magna Straus (300 ind./L) or Moina macrocopa Straus (500 ind./L), The growth of M. aeruginosa, based on cell number, was also significantly different from populations cultured in the media with and without filtered zooplankton water from the exponential growth phase. In the 6-day experiment, the growth pattern of M. aeruginosa cultured with ZCMF was clearly different to control with-out ZCMF. Mean number of cells/particle and particle bio-volume of M. aeruginosa increased significantly from the day 2 for the Daphnia-CMF or Moina-CMF treat-ments. Microcystin production was promoted showing from 18.7 to 55 ${\mu}g/g$-dry cell in the zooplankton treatments relative to the controls. At peaked level on day 4, the highest level of up to $70.5{\pm}16.8\;{\mu}g/g$-dry cell was observed in the D. magna treatment. This study suggested that 'info-chemicals' from zooplankton might induce the increase of algal growth rates, colony formation and microcystin production, these seem to be advantageous to the alga and thus as a grazing defense mechanism.

Effects of Nonylphenol and Diethylhexyl Phthalate on the Population Growth of Freshwater HABs, Microcystis and Stephanodiscus (내분비교란물질이 조류의 성장에 미치는 영향)

  • Lee, Ju-Han;Kim, Baik-Ho;Han, Myung-Soo
    • Korean Journal of Ecology and Environment
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    • v.38 no.3 s.113
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    • pp.304-312
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    • 2005
  • Effects of two endocrine disrupters (EDs) , nonylphenol (NP) and diethylhexyl phthalate (DEHP), on the population growth and morphology of two freshwater HABs (harmful algal blooms) , Microcystis aeruginosa and Stephanodiscus hantzschii, which frequently evoked the hazard bloom in an eutrophic lakes and reservoirs worldwide, were examined with seven different concentrations of EDs (0.01, 0.05, 0.1, 1,2, 2.5 and 3 ppm). Even at concentration below 0.01 ppm, NP strongly inhibited the algal growth of both M. aeruginosa and S. hantzschii, regardless of the algal growth phase. Morphologically, the algal cell treated with NP gradually lost green color in cytoplasm, became smaller in cell size, and then, was hardly seen in microscopic field. On the other hand, DEHP employed did not affect two algae at all concentrations, and rather stimulated the growth by about 10% with a treatment at 3.00 ppm compared to control. These results indicate that the continuous input of EDs, DEHP into the natural water system plays a crucial role to enhance or help an outbreak of algal bloom in eutrophic waters.

Isolation and Genetic Transformation of Primordial Germ Cell (PGC)-Derived Cells from Cattle, Goats, Rabbits and Rats

  • Lee, C.K.;Moore, K.;Scales, N.;Westhusin, M.;Newton, G.;Im, K.S.;Piedrahita, J.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.5
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    • pp.587-594
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    • 2000
  • At present embryonic stem (ES) cells with confirmed pluripotential properties are only available in the mouse. Recently, we were able to isolate, culture and genetically transform primordial germ cell (PGC)-derived cells from pig embryos and demonstrate their ability to contribute to chimera development in the pig. In order to determine whether the system we developed could be used to isolate embryonic germ (EG) cells from other mammalian species, we placed isolated PGCs from cattle, goats, rabbits and rats in culture. Briefly, PGCs were isolated from fetuses of cow (day 30-50), goat (day 25), rabbit (day 15-18) and rat (day 11-12), and plated on STO feeder cells in Dulbecco's modified Eagle's medium (DMEM): Ham's F10 medium (1:1) supplemented with 0.01 mM nonessential amino acids, 2 mM L-glutamine, 0.1 mM $\beta$ - mercaptoethnol, soluble recombinant human stem cell factor (SCF; 40ng/ml), human basic fibroblast growth factor (bFGF; 20ng/ml) and human leukemia inhibitory factor (LIF; 20ng/ml). For maintenance of the cells, colonies were passed to fresh feeders every 7-10 days. In all species tested, we were able to obtain and maintain colonies with ES-like morphology. Their developmental potential was tested by alkaline phosphatase (AP) staining and in vitro differentiation assay. For genetic transformation, cells were electroporated with a construct containing the green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) promoter. GFP-expressing colonies were detected in cattle, rabbits and rats. These results suggest that PGC-derived cells from cattle, goats, rabbits and rats can be isolated, cultured, and genetically transformed, and provide the basis for analyzing their developmental potential and their possible use for the precise genetic modification of these species.

A Study on the Properties of Al doped ZnO (AZO) Thin Films Deposited by RF Magnetron Sputtering (RF 마그네트론 스퍼터링으로 증착된 Al이 도핑 된 ZnO (AZO) 박막의 특성에 대한 연구)

  • Yun, Eui-Jung;Jung, Myung-Hee;Park, Nho-Kyung
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.47 no.7
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    • pp.8-16
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    • 2010
  • In this paper, we investigated the effects of $O_2$ fraction on the properties of Al-doped ZnO (AZO) thin films prepared by radio frequency (RF) magnetron sputtering. Hall, photoluminescence (PL), and X-ray photoelectron spectroscopy (XPS) measurements revealed that the p-type conductivity was exhibited for AZO films with an $O_2$ fraction of 0.9 while the n-type conductivity was observed for films with $O_2$ fractions in range of 0 - 0.6. PL and XPS also showed that the acceptor-like defects, such as zinc vacancies and oxygen interstitials, increased in films prepared by an $O_2$ fraction of 0.9, resulting in the p-type conductivity in the films. Hall results indicated that AZO films prepared by $O_2$ fractions in range of 0 - 0.6 can be used for electrode layers in the applications of transparent thin film transistor. We concluded from the X-ray diffraction analysis that worse crystallinity with a smaller grain size as well as higher tensile stress was observed in the films prepared by a higher $O_2$ fraction, which is related to incorporation of more oxygen atoms into the films during deposition. The study of atomic force microscope suggested that the smoother surface morphology was observed in films prepared by using $O_2$ fraction, which causes the higher resistivity in those films, as evidenced by Hall measurements.

Effect of Exercise Training on Aging Atrophy in Rat Skeletal Muscle II. Effect of Long Term Weight-Training (흰쥐 골격근의 노화성 위축에 대한 운동훈련의 영향 - II. 장기간에 걸친 체중부하 훈련의 영향 -)

  • Park, Sung-Han;Park, Won-Hark;Lee, Yong-Deok;Kim, Jung-Ki
    • Applied Microscopy
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    • v.25 no.4
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    • pp.26-51
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    • 1995
  • The present study was designed to examine effect of long term weight-training on aging atrophy in the rat skeletal muscle. Male rats of 8, 15, and 24 month old were used. Each age groups included control and weight-training for 5 months by using body press apparatus. The histo- and cytochemical, ultrastructural and stereological changes in aging skeletal muscles of the rat were observed in the present study. During the training period the body weight and muscular weight in all groups except the rectus femoris and the gastrocnemius in young age groups remained constant, but muscular weights were increased in the rectus femoris and the gastrocnemius muscles in young age groups. In trained rat, the volume density of muscle fiber type IIA and IIB were increased, but those of type IIC was decreased. Type I remained constant in 8 and 15 month old age groups, but reduced in the tibialis anterior and the gastrocnemius muscles in the 24 month old groups. Some histotological and ultrastructural changes associated with age were found: numerical increase of cytiplasmic vacuoles, lysosomes, lipofuscins, and irregularity of myofibrils. At 24 month old groups some unusual formation of contraction band and muscle splitting were observed. After weight-training, ultrastructural degenerative changes occured in the type I muscle fiber, such as splitting of muscle fiber, disorganization of myofilaments, swelling of mitochondria, accumulation of many lipid droplets, appearance of many lysosomes and residual bodies and necrotic fibers, in the old age groups. But, in the type II muscle fibers hypertrophy of muscle fiber appeared without any noticible damage as the type I. The activities of $Mg^{++}$ -ATPase decreased with age and this enzyme activities in the trained rat were significantly decreased with age. Activities of the acid phosphatase were increased with age and significantly in the trained rat. In stereological analysis, volume density of the myofibrils and the tubular system were increased, on the other hand there mitochondrial capacity was decreased. These experimental results suggested that old rats are not susceptible to be affected by weight-training as young rats, and that physical capacity of the rats must be considered when old rats are exercised for training.

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Effect of Fucoidan Extracted from Hizikia fusiforme on Intestinal Villi and Salmenolla spp. in Broiler Chicks (톳추출 Fucoidan이 병아리 장내 Villi 및 Salmonella 균주에 미치는 영향)

  • 김창혁;박재인
    • Korean Journal of Poultry Science
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    • v.30 no.1
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    • pp.41-47
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    • 2003
  • This study was conducted to investigate the in vivo and in vivo antibiotic effect of crude fucoidan extracted from Hizikia fusiforme, and to investigate any possible structural changes of broiler chick's intestinal villi by the supplementation of fucoidan. Total 84 broiler chicks were randomly assigned to 7 treatments, control and Salmonella typhimurium infection groups. The broiler chicks was infected with Salmonella typhimurium at third days, and antibiotics, fucoidan, dried Hizikia fusiforme, dried Undaria pinnatifida and yeast cell debris was respectively supplemented for each group. Each treatment had 4 chicks with three replications. Extraction yield of crude fucoidan from Hizikia fusiforme was 5.453%. Antibiotic effect of fucoidan was not detected in vitro, inhibition zone and micoorganism growth test. Weight gains of broiler chicks were tend to higher in fucoidan treatment group and yeast cell significance was not found. In in vivo test, the number of viable Salmonella typhimurium was low in the antibiotics and fucoidan treatment groups. The intestinal villi were short in the fucoidan and marine algae treatment groups. The intestinal villi were densely distributed on the large intestinal wall, but the morphology was not different among treatments.

Down-regulation of SENP1 Expression Increases Apoptosis of Burkitt Lymphoma Cells

  • Huang, Bin-Bin;Gao, Qing-Mei;Liang, Wei;Xiu, Bing;Zhang, Wen-Jun;Liang, Ai-Bin
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2045-2049
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    • 2012
  • Objective: To investigate the effect of down-regulation of Sentrin/SUMO-specific protease 1 (SENP1) expression on the apoptosis of human Burkitt lymphoma cells (Daudi cells) and potential mechanisms. Methods: Short hairpin RNA (shRNA) targeting SENP1 was designed and synthesized and then cloned into a lentiviral vector. A lentiviral packaging plasmid was used to transfect Daudi cells (sh-SENP1-Daudi group). Daudi cells without transfection (Daudi group) and Daudi cells transfected with blank plasmid (sh-NC-Daudi group) served as control groups. Flow cytometry was performed to screen GFP positive cells and semiquantitative PCR and Western blot assays were employed to detect the inference efficiency. The morphology of cells was observed under a microscope before and after transfection. Fluorescence quantitative PCR and Western blot assays were conducted to measure the mRNA and protein expression of apoptosis related molecules (caspase-3, 8 and 9). After treatment with $COCl_2$ for 24 h, the mRNA and protein expression of hypoxia inducible factor -$1{\alpha}$ (HIF-$1{\alpha}$) was determined. Results: Sequencing showed the expression vectors of shRNA targeting SENP1 to be successfully constructed. Following screening of GFP positive cells by FCM, semiqualitative PCR showed the interference efficiency was $79.2{\pm}0.026%$. At 48 h after transfection, the Daudi cells became shrunken, had irregular edges and presented apoptotic bodies. Western blot assay revealed increase in expression of caspase-3, 8 and 9 with prolongation of transfection (P<0.05). Following hypoxia treatment, mRNA expression of HIF-$1{\alpha}$ remained unchanged in three groups (P>0.05) but the protein expression of HIF-$1{\alpha}$ markedly increased (P<0.05). However, in the sh-SENP1-Daudi group, the protein expression of HIF-$1{\alpha}$ remained unchanged Conclusion: SENP1-shRNA can efficiently inhibit SENP1 expression in Daudi cells. SENP1 inhibition may promote cell apoptosis. These findings suggest that SENP1 may serve as an important target in the gene therapy of Burkitts lymphoma.

Studies on the Fishery Biology of Pomfrets, Pampus spp. in the Korean Waters 6. Stock Assessment of Korean pomfret, Pampus echinogaster (한국근해 병어류의 자원생물학적 연구 6. 덕대의 자원해석과 관리)

  • KIM Yong Mun;KANG Yong Joo;PARK Byung Ha;LEE Dong Woo;LEE Joo Hee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.22 no.5
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    • pp.306-316
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    • 1989
  • We examined the state of Korean pomfret(Pampus echinogaster) in Korean waters and considered the management strategy of the stock based on the theory yield Per recruit. It is not facile to discriminate Korean pomfret distributed in Korean waters from silver pomfret (p. argenteus) due to the similarities in their external morphologies. For this rea- son, Korean pomfret has been treated in silver pomfret in fisheries statistics of Korea. In this study, we asserted Korean pomfret from pomfrets caught commercially by the morphology, from which we recognized that Korean pomfret took $60\~70\%$ in catch(in weight) and that the smaller the body length, the higher the proportion of Korean pomfret. Parameters estimated for Korean pomfret were as follows: natural mortality(M) = 0.6, fishing mortality(F) =0.924(mean value for $1986\~1988$), age at recruit to fishery($t_r$) =0.19 yrs, age at first capture($t_c$) : 0.49 yrs, and the rate of recruit of age-0 fish to fishery(Q) = 0.29. The results obtained from the theory of yield per recruit indicated that the present state of stock was below the optimum level of exploitation and that the control of fishing intensity rarely had an effect on the increasing of yield. Accordingly, we conclude that proper management can be made by increasing the current age of 0.49 yrs at first capture to 1.5 yrs.

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